A Redox Regulatory System Critical for Mycobacterial Survival in Macrophages and Biofilm Development

English version České info

Nearly one-third of the world’s population is infected with Mycobacterium tuberculosis (Mtb), the causative agent of TB. A key factor that contributes to the widespread infection of Mtb is its capacity to survive inside the host macrophage. Understanding how Mtb withstands the hostile intracellular environment of this phagocytic cell may reveal targets for development of therapeutics that enhance the innate anti-Mtb activities of the macrophage. We discovered a novel signaling pathway in mycobacteria which regulates cellular redox homeostasis through NADH and FAD, regulators of metabolism and redox balance. NADH induces the expression of a protein kinase, PknG, which then phosphorylates the ribosomal protein L13 and promotes its presence in the cytoplasm. L13 therein forms a complex with RenU, a Nudix (Nucleoside diphosphate linked moiety X) hydrolase that degrades NADH and FAD. Genetic disruption of this signaling cascade leads to cellular accumulation of these molecules, increased mycobacterial sensitivity to oxidative stress, impaired surface biofilm growth, and most importantly, reduced survival of Mtb in macrophages.

Vyšlo v časopise: A Redox Regulatory System Critical for Mycobacterial Survival in Macrophages and Biofilm Development. PLoS Pathog 11(4): e32767. doi:10.1371/journal.ppat.1004839
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004839

Souhrn

Zdroje

1. Armstrong JA, Hart DPA (1971) Response of cultured macrophages to Mycobacterium tuberculosis, with observations on fusion of lysosomes with phagosomes. J Exp Med 134 : 713–740. 15776571

2. Russell DG (2001) Mycobacterium tuberculosis: here today, and here tomorrow. Nat Rev Mol Cell Biol 2 : 569–577. 11483990

3. Walburger A, Koul A, Ferrari G, Nguyen L, Prescianotto-Baschong C, et al. (2004) Protein kinase G from pathogenic mycobacteria promotes survival within macrophages. Science 304 : 1800–1804. 15155913

4. Wolff KA, Nguyen HT, Cartabuke RH, Singh A, Ogwang S, et al. (2009) Protein kinase G is required for intrinsic antibiotic resistance in mycobacteria. Antimicrob Agents Chemother 53 : 3515–3519. doi: 10.1128/AAC.00012-09 19528288

5. van der Woude AD, Stoop EJ, Stiess M, Wang S, Ummels R, et al. (2014) Analysis of SecA2-dependent substrates in Mycobacterium marinum identifies protein kinase G (PknG) as a virulence effector. Cell Microbiol 16 : 280–295. doi: 10.1111/cmi.12221 24119166

6. Costerton JW, Stewart PS, Greenberg EP (1999) Bacterial biofilms: a common cause of persistent infections. Science 284 : 1318–1322. 10334980

7. Gefen O, Balaban NQ (2009) The importance of being persistent: heterogeneity of bacterial populations under antibiotic stress. FEMS Microbiol Rev 33 : 704–717. doi: 10.1111/j.1574-6976.2008.00156.x 19207742

8. Kolter R, Losick R (1998) One for all and all for one. Science 280 : 226–227. 9565532

9. Lewis K (2007) Persister cells, dormancy and infectious disease. Nat Rev Microbiol 5 : 48–56. 17143318

10. Ojha A, Anand M, Bhatt A, Kremer L, Jacobs WR Jr., et al. (2005) GroEL1: a dedicated chaperone involved in mycolic acid biosynthesis during biofilm formation in mycobacteria. Cell 123 : 861–873. 16325580

11. Ojha AK, Baughn AD, Sambandan D, Hsu T, Trivelli X, et al. (2008) Growth of Mycobacterium tuberculosis biofilms containing free mycolic acids and harbouring drug-tolerant bacteria. Mol Microbiol 69 : 164–174. doi: 10.1111/j.1365-2958.2008.06274.x 18466296

12. Ojha A, Hatfull GF (2007) The role of iron in Mycobacterium smegmatis biofilm formation: the exochelin siderophore is essential in limiting iron conditions for biofilm formation but not for planktonic growth. Mol Microbiol 66 : 468–483. 17854402

13. Nguyen L, Walburger A, Houben E, Koul A, Muller S, et al. (2005) Role of protein kinase G in growth and glutamine metabolism of Mycobacterium bovis BCG. J Bacteriol 187 : 5852–5856. 16077135

14. Cowley S, Ko M, Pick N, Chow R, Downing KJ, et al. (2004) The Mycobacterium tuberculosis protein serine/threonine kinase PknG is linked to cellular glutamate/glutamine levels and is important for growth in vivo. Mol Microbiol 52 : 1691–1702. 15186418

15. Scherr N, Honnappa S, Kunz G, Mueller P, Jayachandran R, et al. (2007) Structural basis for the specific inhibition of protein kinase G, a virulence factor of Mycobacterium tuberculosis. Proc Natl Acad Sci U S A 104 : 12151–12156. 17616581

16. Bessman MJ, Frick DN, O'Handley SF (1996) The MutT proteins or "Nudix" hydrolases, a family of versatile, widely distributed, "housecleaning" enzymes. J Biol Chem 271 : 25059–25062. 8810257

17. Dos Vultos T, Blazquez J, Rauzier J, Matic I, Gicquel B (2006) Identification of Nudix hydrolase family members with an antimutator role in Mycobacterium tuberculosis and Mycobacterium smegmatis. J Bacteriol 188 : 3159–3161. 16585780

18. Kloosterman H, Vrijbloed JW, Dijkhuizen L (2002) Molecular, biochemical, and functional characterization of a Nudix hydrolase protein that stimulates the activity of a nicotinoprotein alcohol dehydrogenase. J Biol Chem 277 : 34785–34792. 12089158

19. Sassetti CM, Boyd DH, Rubin EJ (2003) Genes required for mycobacterial growth defined by high density mutagenesis. Mol Microbiol 48 : 77–84. 12657046

20. Houben EN, Walburger A, Ferrari G, Nguyen L, Thompson CJ, et al. (2009) Differential expression of a virulence factor in pathogenic and non-pathogenic mycobacteria. Mol Microbiol 72 : 41–52. doi: 10.1111/j.1365-2958.2009.06612.x 19210624

21. Brekasis D, Paget MS (2003) A novel sensor of NADH/NAD+ redox poise in Streptomyces coelicolor A3(2). The EMBO journal 22 : 4856–4865. 12970197

22. Gyan S, Shiohira Y, Sato I, Takeuchi M, Sato T (2006) Regulatory loop between redox sensing of the NADH/NAD(+) ratio by Rex (YdiH) and oxidation of NADH by NADH dehydrogenase Ndh in Bacillus subtilis. Journal of bacteriology 188 : 7062–7071. 17015645

23. Bitoun JP, Liao S, Yao X, Xie GG, Wen ZT (2012) The Redox-Sensing Regulator Rex Modulates Central Carbon Metabolism, Stress Tolerance Response and Biofilm Formation by Streptococcus mutans. PloS one 7: e44766. doi: 10.1371/journal.pone.0044766 23028612

24. Bitoun JP, Nguyen AH, Fan Y, Burne RA, Wen ZT (2011) Transcriptional repressor Rex is involved in regulation of oxidative stress response and biofilm formation by Streptococcus mutans. FEMS microbiology letters 320 : 110–117. doi: 10.1111/j.1574-6968.2011.02293.x 21521360

25. Hull RV, Conger PS 3rd, Hoobler RJ (2001) Conformation of NADH studied by fluorescence excitation transfer spectroscopy. Biophysical chemistry 90 : 9–16. 11321678

26. Mazumder B, Sampath P, Seshadri V, Maitra RK, DiCorleto PE, et al. (2003) Regulated release of L13a from the 60S ribosomal subunit as a mechanism of transcript-specific translational control. Cell 115 : 187–198. 14567916

27. Stuhrmann HB, Koch MH, Parfait R, Haas J, Ibel K, et al. (1977) Shape of the 50S subunit of Escherichia coli ribosomes. Proc Natl Acad Sci U S A 74 : 2316–2320. 329279

28. Chaudhuri S, Vyas K, Kapasi P, Komar AA, Dinman JD, et al. (2007) Human ribosomal protein L13a is dispensable for canonical ribosome function but indispensable for efficient rRNA methylation. RNA 13 : 2224–2237. 17921318

29. Torres M, Condon C, Balada JM, Squires C, Squires CL (2001) Ribosomal protein S4 is a transcription factor with properties remarkably similar to NusA, a protein involved in both non-ribosomal and ribosomal RNA antitermination. The EMBO journal 20 : 3811–3820. 11447122

30. Scott JM, Ju J, Mitchell T, Haldenwang WG (2000) The Bacillus subtilis GTP binding protein obg and regulators of the sigma(B) stress response transcription factor cofractionate with ribosomes. J Bacteriol 182 : 2771–2777. 10781545

31. Mukhopadhyay R, Ray PS, Arif A, Brady AK, Kinter M, et al. (2008) DAPK-ZIPK-L13a axis constitutes a negative-feedback module regulating inflammatory gene expression. Molecular cell 32 : 371–382. doi: 10.1016/j.molcel.2008.09.019 18995835

32. Ray PS, Arif A, Fox PL (2007) Macromolecular complexes as depots for releasable regulatory proteins. Trends in biochemical sciences 32 : 158–164. 17321138

33. Warner JR, McIntosh KB (2009) How common are extraribosomal functions of ribosomal proteins? Molecular cell 34 : 3–11. doi: 10.1016/j.molcel.2009.03.006 19362532

34. Niebisch A, Kabus A, Schultz C, Weil B, Bott M (2006) Corynebacterial protein kinase G controls 2-oxoglutarate dehydrogenase activity via the phosphorylation status of the OdhI protein. J Biol Chem 281 : 12300–12307. 16522631

35. O'Hare HM, Duran R, Cervenansky C, Bellinzoni M, Wehenkel AM, et al. (2008) Regulation of glutamate metabolism by protein kinases in mycobacteria. Mol Microbiol 70 : 1408–1423. doi: 10.1111/j.1365-2958.2008.06489.x 19019160

36. Tretter L, Adam-Vizi V (2005) Alpha-ketoglutarate dehydrogenase: a target and generator of oxidative stress. Philosophical transactions of the Royal Society of London Series B, Biological sciences 360 : 2335–2345. 16321804

37. Kohanski MA, Dwyer DJ, Hayete B, Lawrence CA, Collins JJ (2007) A common mechanism of cellular death induced by bactericidal antibiotics. Cell 130 : 797–810. 17803904

38. Smith T, Wolff KA, Nguyen L (2013) Molecular biology of drug resistance in Mycobacterium tuberculosis. Curr Top Microbiol Immunol 374 : 53–80. doi: 10.1007/82_2012_279 23179675

39. Kolodkin-Gal I, Elsholz AK, Muth C, Girguis PR, Kolter R, et al. (2013) Respiration control of multicellularity in Bacillus subtilis by a complex of the cytochrome chain with a membrane-embedded histidine kinase. Genes Dev 27 : 887–899. doi: 10.1101/gad.215244.113 23599347

40. Dietrich LE, Teal TK, Price-Whelan A, Newman DK (2008) Redox-active antibiotics control gene expression and community behavior in divergent bacteria. Science 321 : 1203–1206. doi: 10.1126/science.1160619 18755976

41. Depas WH, Hufnagel DA, Lee JS, Blanco LP, Bernstein HC, et al. (2013) Iron induces bimodal population development by Escherichia coli. Proc Natl Acad Sci U S A 110 : 2629–2634. doi: 10.1073/pnas.1218703110 23359678

42. Singh R, Mailloux RJ, Puiseux-Dao S, Appanna VD (2007) Oxidative stress evokes a metabolic adaptation that favors increased NADPH synthesis and decreased NADH production in Pseudomonas fluorescens. J Bacteriol 189 : 6665–6675. 17573472

43. Geier H, Mostowy S, Cangelosi GA, Behr MA, Ford TE (2008) Autoinducer-2 triggers the oxidative stress response in Mycobacterium avium, leading to biofilm formation. Appl Environ Microbiol 74 : 1798–1804. doi: 10.1128/AEM.02066-07 18245256

44. Demple B (2008) Community organizers and (bio)filmmaking. Nature chemical biology 4 : 653–654. doi: 10.1038/nchembio1108-653 18936747